A recombinant antigen with potential for serodiagnosis of Echinococcus granulosus infection in dogs.
Antibodies specific for Echinococcus granulosus were affinity purified from dog serum on transfer blots containing putative serodiagnostic antigens. These antibodies and serum pools derived from dogs with E. granulosus infections were used to screen a lambda gt11 cDNA library constructed using E. granulosus protoscolex mRNA. Nine definitive antigenic clones were isolated and characterized, of which one (c10P1) gave strong specific reactions in plaque immunoassay with sera from E. granulosus infected dogs. These clones were all subcloned into the plasmid vector pGEX-1. Antigenicity of clones was confirmed in colony immunoassay and/or immunoblot. Glutathione S-transferase (GST) fusion proteins of individual subclones were produced in Escherichia coli, purified by affinity chromatography and evaluated in ELISA using sera from dogs with infections of E. granulosus, Taenia spp. or nematodes, and helminth-free dog sera. The GST fusion protein 10P1 showed a specificity of 100% in ELISA for diagnosis of E. granulosus infection in dogs despite its relatively low sensitivity. Further investigations aim to identify additional recombinant antigens and test 10P1 expressed in alternative expression systems to increase diagnostic sensitivity of the ELISA.
Gasser
R B
RB
University of Melbourne, Werribee, Victoria, Australia.
Lightowlers
M W
MW
Rickard
M D
MD
D000818:Animals; D000909:Antibodies, Helminth / Q000032:analysis*; D000947:Antigens, Helminth / Q000276:immunology*; D004283:Dog Diseases / Q000175:diagnosis*; D004285:Dogs; D004443:Echinococcosis / Q000175:diagnosis / Q000662:veterinary*; D004446:Echinococcus / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D011994:Recombinant Proteins / Q000276:immunology
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D011994:Recombinant Proteins